Codon-Optimized CRISPR Base Editors for Plant Genomic Editing
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current base editing tools for plants have low editing efficiency across various species, with rice being the only exception showing variable results, while maize and wheat exhibit very low editing frequencies, limiting the applicability of precise genomic alterations in plants.
Innovation Solution
Development of codon-optimized CRISPR-Cas nucleases and nucleic acid constructs that include a deaminase domain, specifically designed for plant expression, to enhance the efficiency of base editing by optimizing the nucleic acid sequences and incorporating promoter regions with introns, thereby improving the expression and activity of base editors in plants.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If base editing tools are used in plants, then genomic modifications can be introduced, but editing efficiency is low across most plant species
Solution Approach 1:
The patent applies codon optimization to change the nucleotide sequence parameters of the base editor genes without altering the amino acid sequence. This parameter change optimizes translation efficiency in plant systems, directly addressing the low editing efficiency problem while maintaining the desired protein function and reliability
Solution Approach 2:
The patent introduces species-specific codon optimization strategies, applying different codon usage patterns tailored to specific plant species. This local quality approach ensures that each plant species receives optimized sequences matched to its specific translational machinery, improving editing efficiency without compromising reliability across diverse species
2Adaptability or versatility
If Cas9 gene editing is used, then mutations can be introduced at targeted locations, but the mutations are unpredictable and typically insertions or deletions
Solution Approach 1:
The patent uses base editors as an intermediary tool between traditional Cas9 cutting and desired genomic alterations. Instead of relying on unpredictable indels from double-strand breaks, the base editor directly converts specific bases (C→T or A→G) through deamination, providing predictable and precise sequence changes while maintaining the ability to target specific genomic locations
Solution Approach 2:
The patent replaces the mechanical cutting and error-prone repair mechanism of Cas9 with a chemical modification mechanism. The base editor uses deaminase domains to chemically convert bases directly, substituting the unpredictable biological repair process with a controlled chemical reaction that yields predictable outcomes
Data Source
AI summary
This invention relates to CRISPR-Cas nucleases codon optimized for expression in plants and nucleic acid constructs encoding base editors comprising a CRISPR-Cas nuclease and a deaminase domain, wherein the nucleic acid constructs are optimized for expression in a plant. The invention further relates to methods of modifying nucleic acids using the nucleic acid constructs.